| Ribonuclease H specifically degrades only the RNA strand in RNA-DNA hybrids. It does not hydrolyze the phosphodiester bonds within single-stranded or double-stranded DNA and RNA. |
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| Catalog # | R2021 |
| Unit Definition | One unit catalyzes the formation of 1nmol of acid soluble products in 20min at 37ºC. Enzyme activity is assayed in the following mixture: 20mM Tris-HCl (H 7.8), 40 mM KCl, 8mM MgCl2, 1mM DTT, 24uM [3H]-POLY(A)-POLY(dT), 0.03mg/ml BSA AND 4% Glycerol. |
| 10X Reaction buffer (included) | R2021A |
| Supplied as a liquid in 200mM Tris HCl, pH 8.0, 400mM KCl, 80mM MgCl2, 10mM DTT. |
| Applications | Suitable for use in Removal of mRNA prior to the synthesis of the 2nd strand of cDNA, Removal of mRNA poly(A) sequences after hybridization with oligo(dT), Site-specific cleavage of RNA and Studies of in vitro polyadenylation rection products. |
| Inhibition and Inactivation | Inhibitors: metal chelators, SH-blocking reagents. |
| -Inactivated by heating at 65ºC for 10min. |
| Endodeoxyribonuclease Assay | No detectable conversion of covalently closed circular DNA to nicked DNA was observed after incubation of 10 units of Ribonuclease H with 1ug of pBR322 DNA |
| in 50ul of reaction buffer for 1 hour at 37°C. |
| Exodeoxyribonuclease Assay | 0% of the total radioactivity was released into trichloroacetic acid-soluble fraction after incubation of 10 units of Ribonuclease H with 1ug of sonicated |
| E.coli [3H]-DNA in 50ul of reaction buffer for 1 hour at 37°C. |
| Ribonuclease Assay | 0% of the total radioactivity was released into trichloroacetic acid-soluble fraction after incubation of 10 units of Ribonuclease H with 1ug of [3H]-RNA in 50ul of reaction buffer for 1 hour at 37°C. |
| Storage and Stability | For long-term storage, aliquot and store at -20°C. Aliquots are stable for at least 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. |
| CAS Number | 9050-76-4 |
| Molecular Weight | 19kD |
| Source | E. coli MRE-600 cells |
| Concentration | ~5u/ul |
| Form | Supplied as a liquid in 25mM HEPES-KOH, pH 8.0, 50mM KCl, 1mM DTT, 1mM EDTA, 0.1mg/ml BSA, and 50% glycerol. |
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| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
| Alternate names | E. coli; cRNAse H |
